SERPINH1 Antibody

What is SERPINH1 and what cellular functions does it perform?

SERPINH1 (Serpin Family H Member 1), also known as HSP47, is a member of the serpin superfamily of serine proteinase inhibitors. It functions as a collagen-specific molecular chaperone localized in the endoplasmic reticulum. The protein plays a critical role in collagen biosynthesis through:

• Binding specifically to collagen molecules

• Facilitating proper folding of procollagen

• Supporting the structural assembly and maturation of collagen

• Preventing premature aggregation of collagen molecules

SERPINH1 has a calculated and observed molecular weight of approximately 46 kDa . The gene encoding SERPINH1 is located on chromosome 11, and its expression can be induced by heat shock, reflecting its role as a heat shock protein .

What applications are validated for SERPINH1 antibodies in research settings?

SERPINH1 antibodies have been validated for multiple applications in research, with specific recommended protocols:

For IHC applications, it's recommended to perform antigen retrieval with TE buffer pH 9.0 or alternatively with citrate buffer pH 6.0 . Research consistently shows that antibody titration should be performed for each experimental system to achieve optimal results .

How can researchers validate SERPINH1 antibody specificity?

Ensuring antibody specificity is crucial for obtaining reliable experimental results. For SERPINH1 antibodies, several validation methods have been established:

• RNAi knockdown validation: Confirming reduced signal in SERPINH1 siRNA-treated samples compared to control siRNA

• Multi-application testing: Verifying consistent detection across different applications (WB, IHC, IF, FC)

• Positive control verification: Testing in cell lines and tissues with known SERPINH1 expression (e.g., NIH-3T3, A431, HeLa, HepG2)

• Molecular weight confirmation: Ensuring detection at the expected 46 kDa size

• Knockout validation: Using SERPINH1 knockout samples as negative controls

Published studies have successfully employed these validation approaches, with particularly strong evidence from knockdown experiments demonstrating antibody specificity .

How does SERPINH1 expression correlate with cancer prognosis?

Analysis across multiple cancer types reveals that SERPINH1 serves as a significant prognostic biomarker:

Research using Cox regression analysis has confirmed that SERPINH1 functions as an independent prognostic factor in these cancer types . Studies have demonstrated that knocking down SERPINH1 significantly inhibits tumor cell invasion, migration, and proliferation, further supporting its role in cancer progression .

What mechanisms explain SERPINH1's influence on tumor immune microenvironment?

SERPINH1 exerts complex effects on the tumor immune microenvironment through several mechanisms:

• Immune cell infiltration influence:

  • Negatively correlates with CD8+ T cells infiltration

  • Positively associates with M2 macrophage infiltration

  • These relationships suggest SERPINH1 may promote immunosuppression by inhibiting CD8+ T cell function while enhancing M2 macrophage recruitment

• Immune checkpoint interaction:

  • Strongly correlates with expression of immune-inhibiting genes including CD276, TGFBI, VEGFA, HAVCR2, IL10, and ADORA2A

  • Positively associates with immunoactivating genes ENTPD1, ICAM1, and TNFRSF4

• Chemokine regulation:

  • Close connection with specific chemokines, particularly CCL26 and CCL11

  • Related to chemokine receptors including CXCR4, CCR1, and CCR10

Single-cell RNA sequencing data revealed higher SERPINH1 expression in tumor cells and endothelial cells compared to immune cells in kidney renal clear cell carcinoma, suggesting cell-type-specific functions .

How might SERPINH1 serve as a target for cancer immunotherapy strategies?

SERPINH1 shows promising potential as an immunotherapy target based on several research findings:

• Immunotherapy response prediction: Lower SERPINH1 expression correlates with better response to immune checkpoint blockade (ICB) therapy

• SERPINH1-related score: Effective method for identifying patients likely to respond to immunotherapy and chemotherapy, particularly in osteosarcoma

• Molecular targeting potential: Given that SERPINH1 expression is significantly higher in tumor cells than immune cells, targeting it may preferentially affect malignant cells

• Immune checkpoint association: Strong correlation with multiple immune checkpoint genes suggests SERPINH1 may modulate response to checkpoint inhibitors

TIDE (Tumor Immune Dysfunction and Exclusion) analysis has prioritized SERPINH1 for mechanistic follow-up research in immunotherapy contexts . The protein's role in collagen biosynthesis and extracellular matrix remodeling provides additional mechanisms through which SERPINH1-targeted therapies might modify the tumor microenvironment to enhance anti-tumor immunity .

What are the optimal conditions for Western blot detection of SERPINH1?

For optimal Western blot detection of SERPINH1, researchers should consider the following protocol guidelines:

• Sample preparation: Effective protein extraction from HT-1080, A2780, A431, HeLa, NIH/3T3 cells as validated positive controls

• Antibody dilution: Primary antibody at 1:1000-1:6000 dilution depending on specific antibody clone and sample type

• Expected band: SERPINH1 is detected at approximately 46 kDa

• Blocking solution: Typically 5% non-fat milk or BSA in TBST

• Incubation conditions: Primary antibody incubation overnight at 4°C yields optimal results

• Signal detection: Both chemiluminescence and fluorescence-based detection systems are suitable

Publications using SERPINH1 antibodies for Western blot have successfully employed these conditions to detect endogenous protein levels across multiple human and mouse cell lines .

How should researchers optimize immunofluorescence protocols for SERPINH1?

For successful immunofluorescence detection of SERPINH1, follow these methodological considerations:

• Fixation: 4% paraformaldehyde for 15-20 minutes at room temperature

• Permeabilization: 0.1-0.5% Triton X-100 in PBS for 5-10 minutes

• Blocking: 5-10% normal serum (matched to secondary antibody host) with 1% BSA

• Primary antibody: Apply at 1:200-1:800 dilution; for frozen sections use 1:400-1:1600

• Incubation: Overnight at 4°C or 1-2 hours at room temperature

• Secondary antibody: Fluorophore-conjugated antibody at manufacturer's recommended dilution

• Counterstaining: DAPI for nuclear visualization

• Mounting: Anti-fade mounting medium to prevent photobleaching

HeLa cells serve as reliable positive controls for IF/ICC applications, with SERPINH1 primarily localized to the endoplasmic reticulum . Confocal microscopy is particularly effective for visualizing the characteristic reticular pattern of this ER-resident protein.

What strategies help troubleshoot common issues with SERPINH1 antibody applications?

When encountering difficulties with SERPINH1 antibody experiments, consider these troubleshooting approaches:

For validated detection in difficult tissues, antigen retrieval with TE buffer pH 9.0 is recommended, with citrate buffer pH 6.0 as an alternative . Titration of the antibody is essential for each experimental system to achieve optimal signal-to-noise ratio .

What mechanisms link SERPINH1 to cancer progression and metastasis?

SERPINH1 contributes to cancer progression through multiple interconnected mechanisms:

• Extracellular matrix remodeling:

  • Promotes procollagen maturation critical for tumor metastasis

  • Interacts with various collagens in the protein-protein interaction network

  • Facilitates structural changes in tumor microenvironment

• Cellular invasion pathways:

  • Single-cell analysis reveals SERPINH1 association with invasion, DNA repair, inflammation, metastasis, and angiogenesis in glioblastoma

  • Strongly correlates with epithelial-mesenchymal transition (EMT) across multiple tumor types

• Cell proliferation regulation:

  • SERPINH1 overexpression promotes proliferation in human retinal endothelial cells (HRECs)

  • Knockdown experiments demonstrate reduced proliferation in multiple cancer cell types

• Molecular targeting:

  • SERPINH1 has been identified as a target of miR-29b

  • miR-29b suppresses SERPINH1 expression in HRECs

  • This regulatory mechanism may be exploited for therapeutic intervention

Gene set enrichment analysis (GSEA) has confirmed that SERPINH1 overexpression is related to malignant progression of tumors across multiple cancer types .

How is SERPINH1 implicated in immune-related conditions beyond cancer?

SERPINH1's biological roles extend to several immune-related conditions:

• Rheumatoid arthritis:

  • Autoantibodies to SERPINH1 have been found in patients with rheumatoid arthritis

  • May serve as an autoantigen in autoimmune joint disease

• Fibrotic disorders:

  • Critical regulator of collagen biosynthesis and structural assembly

  • Inhibition of SERPINH1 or its interaction with procollagen reduces procollagen secretion from fibroblasts

  • Potential therapeutic target for management of fibrosis

• Diabetic retinopathy:

  • SERPINH1 overexpression promotes human retinal endothelial cell proliferation and migration

  • Targeted by miR-29b, suggesting a regulatory pathway that can be manipulated

  • Identified as a hub gene in diabetic retinal tissues through bioinformatics analysis

The connection between SERPINH1 and these conditions underscores its importance as a potential therapeutic target beyond oncology, particularly in diseases characterized by dysregulated collagen production or autoimmune responses .

How does SERPINH1 expression influence chemotherapy response?

Emerging research indicates that SERPINH1 may impact response to chemotherapy in several ways:

• Predictive biomarker potential:

  • The SERPINH1-related score effectively identifies patients likely to respond to both immunotherapy and chemotherapy, particularly in osteosarcoma

  • Expression levels correlate with treatment outcomes in multiple cancer types

• Drug resistance mechanisms:

  • SERPINH1's role in collagen biosynthesis may contribute to extracellular matrix remodeling that affects drug penetration

  • Association with DNA repair pathways suggests potential influence on DNA-damaging chemotherapeutics

• Tumor microenvironment modulation:

  • By affecting immune cell infiltration and function, SERPINH1 may indirectly influence chemotherapy efficacy

  • Immunosuppressive environment associated with high SERPINH1 expression could reduce the effectiveness of certain chemotherapeutic agents

• Combined treatment approach:

  • Research suggests potential for targeting SERPINH1 to enhance chemotherapy response

  • Strategy of combining SERPINH1 inhibition with conventional chemotherapy warrants further investigation

While research specifically examining SERPINH1's impact on chemotherapy response is still emerging, the protein's roles in cancer cell survival, proliferation, and tumor microenvironment modulation strongly suggest its relevance to treatment resistance mechanisms .

What emerging technologies are advancing SERPINH1 research?

Several cutting-edge technologies are enhancing our understanding of SERPINH1:

• Single-cell RNA sequencing (scRNA-seq):

  • Reveals cell-type-specific expression patterns of SERPINH1 in the tumor microenvironment

  • Identifies SERPINH1 expression as highest in tumor cells and endothelial cells but lowest in macrophages and mast cells

  • Enables pseudotime trajectory analysis of SERPINH1 function during cell differentiation

• CRISPR/Cas9 gene editing:

  • Facilitates creation of SERPINH1 knockout models for definitive functional studies

  • Provides essential negative controls for antibody validation

• Computational prediction algorithms:

  • TIDE (Tumor Immune Dysfunction and Exclusion) analysis prioritizes SERPINH1 for immunotherapy research

  • CancerSEA exploration of SERPINH1 function at single-cell level reveals associations with invasion, DNA repair, and metastasis

• Multi-omics integration:

  • Combines gene expression data with clinical outcomes and immune infiltration analysis

  • ESTIMATE algorithm correlates SERPINH1 with Immune, Stromal, and ESTIMATE scores to comprehensively assess tumor microenvironment

These technologies collectively enable more precise characterization of SERPINH1's roles in normal physiology and disease pathogenesis .

What are the key methodological considerations for studying SERPINH1 in primary patient samples?

When investigating SERPINH1 in clinical specimens, researchers should address these methodological challenges:

These methodological considerations ensure robust and clinically relevant findings when studying SERPINH1 in patient samples .

How might SERPINH1-targeting therapeutic strategies be developed and optimized?

Development of SERPINH1-targeted therapies requires consideration of multiple approaches:

• Small molecule inhibitors:

  • Target SERPINH1's collagen-binding activity

  • Disrupt chaperone function in the endoplasmic reticulum

  • Focus on cancer-specific vulnerabilities created by SERPINH1 dependency

• RNA interference strategies:

  • siRNA or shRNA targeting SERPINH1 has shown efficacy in reducing cancer cell proliferation and invasion

  • Delivery systems must be optimized for specific tissue targeting

  • Can be combined with conventional therapies for enhanced effect

• miRNA-based approaches:

  • miR-29b has been identified as a regulator of SERPINH1

  • miRNA mimics could potentially downregulate SERPINH1 expression

  • Requires tissue-specific delivery systems

• Patient selection biomarkers:

  • SERPINH1 expression levels as predictive biomarkers for therapy response

  • SERPINH1-related scoring systems to identify likely responders

  • Integration with other genomic and immune markers

• Combination therapy strategies:

  • Combine SERPINH1 inhibition with immune checkpoint inhibitors

  • Pair with conventional chemotherapy for synergistic effects

  • Target both SERPINH1 and related pathways for maximum efficacy